Quantitative RT-PCR Analysis of DRAM Gene

Total RNA was extracted from cells and tissues using RNAiso (Takara, Shiga, Japan). Then, RNA was converted to cDNA using PrimeScript RT reagent kit with genomic DNA Eraser (Takara). The quantification of gene expression was determined with the SYBR Green polymerase chain reaction (PCR) kit (Qiagen). The following primers were used for quantitative reverse transcription PCR (qRT-PCR) analysis in this study: DRAM (mouse): 5′-CAGCCTTCATCATCTCCTACG-3′ and 5′-ATGCAGAGAAGTTTATCATG-3′ (27 (link)), DRAM (human): 5′-TCAAATATCACCATTGATTTCTGT-3′ and 5′-GCCACATACGGATGGTCATCTCTG-3′ (16 (link)), β-actin (mouse): 5′-CAGCTTCTTTGCAGCTCCTT-3′ and 5′-CACGATGGAGGGAATACAG-3′, and β-actin (human): 5′-CATCCTCACCCTGAAGTACCCC-3′ and 5′-AGCCTGGATGCAA CGTACATG-3′.